{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T06:26:48Z","timestamp":1792132008616,"version":"4.4.0"},"reference-count":34,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,12,15]],"date-time":"2024-12-15T00:00:00Z","timestamp":1734220800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,12,15]],"date-time":"2024-12-15T00:00:00Z","timestamp":1734220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["3\u20103249\/Z\/16\/Z"],"award-info":[{"award-number":["3\u20103249\/Z\/16\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["3\u20133249\/Z\/16\/Z"],"award-info":[{"award-number":["3\u20133249\/Z\/16\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007849","name":"British Antarctic Survey","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007849","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","award":["MR\/K015850\/1"],"award-info":[{"award-number":["MR\/K015850\/1"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","award":["MR\/K02292X\/1"],"award-info":[{"award-number":["MR\/K02292X\/1"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000266","name":"Engineering and Physical Sciences Research Council","doi-asserted-by":"publisher","award":["EP\/H018301\/1"],"award-info":[{"award-number":["EP\/H018301\/1"]}],"id":[{"id":"10.13039\/501100000266","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000266","name":"Engineering and Physical Sciences Research Council","doi-asserted-by":"publisher","award":["EP\/L015889\/1"],"award-info":[{"award-number":["EP\/L015889\/1"]}],"id":[{"id":"10.13039\/501100000266","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The Antarctic seabed harbors significant biodiversity, and almost 90% of oceanic environments are permanently below 5 \u00b0C (i.e., deep sea and polar regions). However, organisms whose entire lifecycle occurs around 0 \u00b0C are understudied, leaving this large and diverse proportion of the global biome poorly understood. To address this question at the cellular level, tools are required for high\u2010resolution imaging of biological systems under physiological conditions. This poses severe technical challenges. High\u2010resolution imaging objectives require short working distances and immersion media, causing rapid heat transfer from the microscope to the sample. This affects the viability of live specimens and the interpretability of results. Here, we present a method for high\u2010fidelity imaging of live biological samples at temperatures of around, or below, 0 \u00b0C. It relies on hardware additions to traditional microscopy, namely as a cooling collar, 10% ethanol as an immersion medium, and nitrogen flow to reduce condensation It can be straightforwardly implemented on different microscopy modalities, including super\u2010resolution imaging. The method is demonstrated in live cell cultures derived from Antarctic fish and highlights the need to maintain physiological conditions for these fragile samples. Future applications include evolutionary biology, biophysics and biotechnology.<\/jats:p>","DOI":"10.1002\/smtd.202401682","type":"journal-article","created":{"date-parts":[[2024,12,16]],"date-time":"2024-12-16T02:40:27Z","timestamp":1734316827000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A High\u2010Resolution Microscopy System for Biological Studies of Cold\u2010Adapted Species Under Physiological Conditions"],"prefix":"10.1002","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-8018-8600","authenticated-orcid":false,"given":"Anne\u2010Pia M.","family":"Marty","sequence":"first","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology University of Cambridge  Phillipa Fawcett Drive Cambridge CB3 0AS UK"},{"name":"British Antarctic Survey  High Cross, Madingley Road Cambridge CB3 0ET UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9078-9716","authenticated-orcid":false,"given":"Edward N.","family":"Ward","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology University of Cambridge  Phillipa Fawcett Drive Cambridge CB3 0AS UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-5804-679X","authenticated-orcid":false,"given":"Jacob R.","family":"Lamb","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology University of Cambridge  Phillipa Fawcett Drive Cambridge CB3 0AS UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9795-2571","authenticated-orcid":false,"given":"Francesca W.","family":"van Tartwijk","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology University of Cambridge  Phillipa Fawcett Drive Cambridge CB3 0AS UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3479-6791","authenticated-orcid":false,"given":"Lloyd S.","family":"Peck","sequence":"additional","affiliation":[{"name":"British Antarctic Survey  High Cross, Madingley Road Cambridge CB3 0ET UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melody S.","family":"Clark","sequence":"additional","affiliation":[{"name":"British Antarctic Survey  High Cross, Madingley Road Cambridge CB3 0ET UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5194-0962","authenticated-orcid":false,"given":"Clemens F.","family":"Kaminski","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology University of Cambridge  Phillipa Fawcett Drive Cambridge CB3 0AS UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,12,15]]},"reference":[{"key":"e_1_2_10_1_1","first-page":"391","volume-title":"Principles of Thermal Ecology: Temperature, Energy, and Life","author":"Clarke A.","year":"2017"},{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1201\/9780429454455"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1017\/S1464793105006871"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2008.02.001"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtherbio.2020.102689"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jpcb.7b06933"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10439-010-9963-z"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2013.05.009"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.2007.01730.x"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2008.12.004"},{"key":"e_1_2_10_11_1","volume":"111","author":"Young L. J.","year":"2016","journal-title":"J. Vis. Exp. JoVE"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.7b00218"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms10980"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.2144\/000112257"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-11024-z"},{"key":"e_1_2_10_16_1","unstructured":"R.Chandler The Anatomy of an Objective Lens https:\/\/www.olympus\u2010lifescience.com\/en\/discovery\/the\u2010anatomy\u2010of\u2010an\u2010objective\u2010lens\/(accessed: May 2024)."},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/jp037222e"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1364\/OE.23.003770"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-physchem-090319-051546"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/cphc.201800131"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jphotochem.2020.112435"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1523\/ENEURO.0138-22.2022"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21666-7"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2448"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2013.05.004"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.0269-8463.2004.00903.x"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00442-007-0858-0"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2006.1947"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0954102092000622"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pocean.2018.09.001"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.156.3772.257"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1098\/rsos.211272"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12985"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1697"}],"container-title":["Small Methods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.202401682","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/smtd.202401682","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.202401682","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T09:09:02Z","timestamp":1774343342000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/smtd.202401682"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,15]]},"references-count":34,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2025,5]]}},"alternative-id":["10.1002\/smtd.202401682"],"URL":"https:\/\/doi.org\/10.1002\/smtd.202401682","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.09.17.613460","asserted-by":"object"}]},"ISSN":["2366-9608","2366-9608"],"issn-type":[{"value":"2366-9608","type":"print"},{"value":"2366-9608","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,15]]},"assertion":[{"value":"2024-10-08","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-12-15","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"2401682"}}